Inhibition of protein degradation increases the Bt protein concentration in Bt cotton

Bacillus thuringiensis(Bt)cotton production is challenged by two main problems,i.e.,the low concentration of Bt protein at the boll setting stage and the lowest insect resistance in bolls among all the cotton plant’s organs.Therefore,increasing the Bt protein concentration at the boll stage,especially in bolls,has become the main goal for increasing insect resistance in cotton.In this study,two protein degradation inhibitors(ethylene diamine tetra acetic acid(EDTA)and leupeptin)were sprayed on the bolls,subtending leaves,and whole cotton plants at the peak flowering stage of two Bt cultivars(medium maturation Sikang 1(SK1))and early maturation Zhongmian 425(ZM425)in 2019 and 2020.The Bt protein content and protein degradation metabolism were assessed.The results showed that the Bt protein concentrations were enhanced by 21.3 to 38.8%and 25.0 to 38.6%in the treated bolls of SK1 and ZM425 respectively,while they were decreased in the subtending leaves of these treated bolls.In the treated leaves,the Bt protein concentrations increased by 7.6 to 23.5%and 11.2 to 14.9%in SK1 and ZM425,respectively.The combined application of EDTA and leupeptin to the whole cotton plant increased the Bt protein concentrations in both bolls and subtending leaves.The Bt protein concentrations in bolls were higher,increasing by 22.5 to 31.0%and 19.6 to 32.5%for SK1 and ZM425,respectively.The organs treated with EDTA or/and leupeptin showed reduced free amino acid contents,protease and peptidase activities and significant enhancements in soluble protein contents.These results indicated that inhibiting protein degradation could improve the protein content,thus increasing the Bt protein concentrations in the bolls or/and leaves of cotton plants.Therefore,the increase in the Bt protein concentration without yield reduction suggested that these two protein degradation inhibitors may be applicable for improving insect resistance in cotton production.

Effects of Dietary Corn Gluten Meal on Growth Performance and Protein Metabolism in Relation to IGF-I and TOR Gene Expression of Juvenile Cobia (Rachycentron canadum)

A growth experiment was conducted on cobia(Rachycentron canadum,initial weight 108.2 g ± 3.0 g) to investigate the effects of dietary corn gluten meal(CGM) levels on the fish growth,whole body composition and protein metabolism in relation to specific gene expression.Five isonitrogenous(crude protein 45%) and isoenergetic(gross energy 20 kJ g 1) practical diets were formulated by replacing 0%(the control),17.5%,35.0%,52.5%,and 70.0% of fish meal(FM) protein with CGM protein.No significant differences were observed in the survival,feed intake(FI),specific growth rate(SGR),feed efficiency(FE) and protein productive value(PPV) among fish fed diets with 0%,17.5%,35.0%,and 52.5% of CGM protein.However,these indices were significantly lower in fish fed the diet with 70.0% of CGM protein than those in fish fed the control diet(P < 0.05).The whole-body crude protein and lipid contents were significantly lower while the whole-body moisture content was significantly higher in fish fed the diet with 70.0% of CGM protein compared with the control group(P < 0.05).When 70.0% of FM protein was replaced by CGM,plasma total protein and cholesterol contents were significantly lower than those in the control group(P < 0.05).Fish fed the diet with 70.0% of CGM protein had significantly lower hepatic insulin-like growth factor I(IGF-I) expression levels than those in the control group(P < 0.05).However,no significant differences were observed in hepatic target of rapamycin(TOR),dorsal muscle IGF-I and TOR expression levels among dietary treatments.Results of the present study indicated that 52.5% of FM protein could be replaced by CGM in the diets without significant influences on the growth,feed utilization and protein metabolism of juvenile cobia.The present results might be useful for developing cost effective and sustainable cobia dietary formulations.

Effects of Chinese herbal medicine on plasma glucose,protein and energy metabolism in sheep

Background： The use of antibiotics in animal diets is facing negative feedback due to the hidden danger of drug residues to human health. Traditional Chinese herbal medicine has been used to replace antibiotics in the past two decades and played an increasingly important role in livestock production. The present study was carried out to assess the feeding effects of a traditional nourishing Chinese herbal medicine mixture on kinetics of plasma glucose, protein and energy metabolism in sheep. Ruminal fermentation characteristics were also determined. Methods： Four sheep were fed on either mixed hay （MH-diet） or MH-diet supplemented with 2% of Chinese herbal medicine （mixture of Astragalus root, Angelica root and Atractylodes rhizome; CHM-diet） over two 3S-day periods using a crossover design. The turnover rate of plasma glucose was measured with an isotope dilution method using [U-^13C]glucose. The rates of plasma leucine turnover and leucine oxidation, whole body protein synthesis （WBPS） and metabolic heat production were measured using the [1-^13C]leucine dilution and open circuit calorimetry. Results： Body weight gain of sheep was higher （P = 0.03） for CHM-diet than for MH-diet. Rumen pH was lower （P = 0.02）, concentration of rumen total volatile fatty acid tended to be higher （P = 0.05） and acetate was higher （P = 0.04） for CHM-diet than for MH-diet. Turnover rates of plasma glucose and leucine did not differ between diets. Oxidation rate of leucine tended to be higher （P = 0.06） for CHM-diet than for MH-diet, but the WBPS did not differ between diets. Metabolic heat production tended to be greater （P = 0.05） for CHM-diet than for MH-diet. Conclusions： The sheep fed on CHM-diet had a higher body weight gain and showed positive impacts on rumen fermentation and energy metabolism without resulting in any adverse response. Therefore, these results suggested that the Chinese herbal medicine mixture should be considered as a potential feed additive for sheep.

Effect of Dietary Folic Acid Supplementation on Growth Performance and Hepatic Protein Metabolism in Early-Weaned Intrauterine Growth Retardation Piglets

To investigate the effects of dietary supplementation with folic acid on growth performance, hepatic protein metabolism and serum biochemical indices of early-weaned intrauterine growth retardation （IUGR） piglets, 24 male （Durocx （LandracexYorkshire）） weaned （14-d-old） IUGR piglets were randomly divided into 3 treatments with 8 replicates of 1 piglet per replicate. The piglets in each treatment were fed basal diet supplementation with either 0 （control）, 5 and 10 mg kg^-1 folic acid. The trial lasted for 21 d. Dietary folic acid supplementation reduced average daily feed intake （ADFI） （P〈0.05）. In addition, the average daily gain （ADG） in 10 mg kg^-1 folic acid group was significantly decreased （P〈0.01） and the ratio of feed：gain （F/G） increased slightly （P〉0.05）. Serum folic acid concentration increased （P〈0.01） with increasing folic acid inclusion, however, serum homocysteine concentration decreased significantly （P〈0.01）. Enhanced serum urine nitrogen （SUN） and diminished serum total protein （TP） as well as liver TP content were observed in 10 mg kg^-1 folic acid group （/＇〈0.05）. Furthermore, the relative mRNA expressions of insulin-like growth factor 1 （IGF-1） and mammalian target of rapamycin （m-TOR） in liver were respectively tended to reduce （P=0.06） and significantly downregulated （P〈0.05） in 10 mg kg1 group, in compared with 5 mg kg1 group. However, when compared with control group, folic acid supplementation had no significant effect on the mRNA abundance of IGF- 1 and m-TOR. The results indicated that supplementation with 10 mg kg-I folic acid impaired growth performance and hepatic protein metabolism of early-weaned IUGR piglets while 5 mg kg-~ folic acid enriched diet exerted limited positive effects.

TUMOR NECROSIS FACTOR－α ALTERS PROTEINMETABOLISM AND CELL－CYCLE KINETICSIN MALIGNANT TUMOR

The effects of tumor necrosis factor-α(TNF) on protein metabolism and cell-cycle kinetics were investigated in malignant tumor. Sprague-Dawley rats, subcutaneously inoculated with Walker 256 carcinosarcoma,were injected intraperitoneally with recombinant human TNF at a dose of 4-75×106 U/kg for 3 consecutive days.Tumor protein metabolism and cell-cycle kinetics were analyzed. The results showed a significant decrease in tumor volume and weight in comparison with control.TNF resulted in significant decrease in tumor Protein fractional synthesis rate, Protein synthesis and fractional growth rate, but no change of tumor protein fractional degradation rate. TNF also resulted in remarkable decline in labelling index and GI phase increase of tumor cells, 6 hours after bromodeoxyuridine injection, by cytometry. The results indicated that TNF inhibits tumor growth as a result of decreases in tumor cell DNA and protein syntheses.

Pesticidal Impact on the Protein Metabolism of Freshwater Field Crab, Oziotelphusa Senex Senex(Fabricius)

The total protein increased in the gills and decreased in the muscle of the freshwater field crab Oziotelphusa senex senex at days 1 and 2 on exposure to lethal concentrations and at days 1 and 10 to sublethal concentrations of furadan, endosulfan, chlorpyrifos, and a mixture of these three in a 100:10: 1 ratio. The increase in the gill protein was greater on exposure to the sublethal concentrations than to the lethal concentrations while the decrease in the muscle protein was greater on exposure to the lethal concentrations than to the sublethal concentrations. In the hepatopancreas, the protein content decreased on exposure to the lethal concentrations, but, in contrast, increased on exposure to the sublethal concentrations. These results clearly indicate that changes in the protein content are not only organ-dependent but also concentration-dependent, i.e., lethal versus sublethal. Irrespective of the changes in the total protein, the levels of free amino acids and the activities of protease, alanine and aspartate aminotransferases, and glutamate dehydrogenase increased in all the three organs of the crabs exposed to the lethal and sublethal concentrations, (more in lethal than in sublethal) and increased at a greater rate over time of exposure. Ammonia toxicity, measured by an increase in the hemolymph ammonia and a decrease in the urea, was also observed at the lethal concentrations of all the three pesticides. The ammonia and urea levels increased in the crabs exposed to the sublethal concentrations. Although the effect of each pesticide on the protein metabolism was similar, the degree of toxicity was the lowest on exposure to furadan, intermediate on exposure to endosulfan and chlorpyrifos, and cumulative on exposure to a mixture of the three pesticides

Changes of carbohydrate and protein metabolism in seedling leaves of a temperature-induced greenable albino mutant line W25 of rice

W25 is a low-temperature-sensitive albino mu-tant line. Temperature not only controls thealbino phenotype expression of W25, but alsodetermines whether it could survive. When thetemperature is lower than 25℃, the leaves ofW25 shows complete albino, but they exhibitsnormal green when temperature is higher than30℃. Meanwhile, at 25℃, it can be greenable

Is intraspecific trait differentiation in Parthenium hysterophorus a consequence of hereditary factors and/or phenotypic plasticity?

Of the various strategies adopted by an invasive plant species for expanding its niche breadth,phenotypic differentiation(either due to plasticity and/or adaptive evolution) is proven to be the most successful.Lately,we studied the persistence of substantial morpho-functional variations within the individuals of alien invasive plant,Parthenium hysterophorus in Chandigarh,India,through field surveys.Based on observed differences,the individuals were categorized into two morphotypes,PAand PB.PAhad higher leaf area,leaf biomass,and chlorophyll content as compared with PB.However,PBhad a higher stem circumference,stem specific density,twig dry matter content,profuse branching,bigger canopy,and better reproductive output than PA.To substantiate the persistence of intraspecific variations in P. hysterophorus and to deduce the possible genesis of these variations,we propagated both the morphotypes under experimental conditions in winter and summer.Apart from the key morpho-functional differences observed during the field studies,protein and carbohydrate metabolism were studied in leaves and roots of the propagated plants.Differences in plant metabolism were observed only during the early growth period,whereas the morpho-functional traits varied in the mature flowering plants.The effect of growth season was highly significant on all the studied morpho-functional and biochemical parameters(p ≤0.05).Parent morphotypes(P) and interactions between morphotypes and seasons significantly affected several growth parameters(p ≤0.05).The analyses revealed that the contrasting growth conditions at the time of transplantation and early growth may regulate the phenotype of P. hysterophorus.The pattern of intraspecific variations observed during the study is justified to consider morphotype PAas winter biotype and morphotype PBas summer biotype of P. hysterophorus.The study points towards the role of plasticity or a combination of genetic and environmental(G×E) factors in producing the phenotypic variability observed in the population of P. hysterophorus.

Precise Calculation System of Total Mix Ration for Lactating Cow

[Objective] The paper was to precisely predict nutrient requirements and optimize ration formula, and explore the inherent feature of ration optimization of dairy cow. [Method] Based on cornell net carbohydrate and protein system （CNCPS） with integrating dynamic prediction models on main nutrient requirements of dairy cattle recommended by NRC （2001） and the CNCPS parameter database accumu- lated by Chinese feed database, the ration nutrient requirement dynamic calculation and total mixed ration （TMR） formula optimizing system for Holstein dairy cow was developed using FOXPRO database system and parametric linear programming algo- rithm. [Result] By optimizing a specific cow ration and analyzing its completed nutri- ents, the results showed that this system could entirely consider a lots of nutrient balances, such as concentrate fraction and forage fraction balance, rumen degrad- able protein and rumen undergradable protein balance, crude protein and lactation net energy balance, fibrous substances （ADF, NDF） and non-fiber carbohydrates （NFC） balance, calcium and phosphate balance, electrolytes balance and trace ele- ment balance etc., and could also calculate intestinal amino acid flow in terms of different models. [Conclusion] By using dynamic mathematical equations and comput- erized program, it can be realized for the ration formula design of lactating cow with all-round interoperable but mutual-constraining each other among ration nutrients.

Creatine Pyruvate Enhances Lipolysis and Protein Synthesis in Broiler Chicken

To assess the effects ofcreatine pyruvate （Cr-Pyr） on lipid and protein metabolism in broiler chickens, a total of 400 1-day-old male birds （Aconred） were randomly allocated to four groups, with each group replicating four times and each replicate involving 25 birds. The broilers were provided with a commercial diet supplemented with Cr-Pyr at 0, 1, 5, or 10% of the diet, respectively, for a period of 3 wk ad libitum （from 22 to 42 d）. In the present study, body weight （BW） and average daily gain （ADG） of broilers decreased in 10% Cr-Pyr group （P〈0.01）, whereas the relative leg and pectoral muscle weights were significantly higher than they were in the control group （P〈0.05）. 5 or 10% Cr-Pyr of diets decreased the abdominal fat rate （AFR, abdominal fat/live weight） of the broilers. The serum or hepatic triglyceride （TG） concentrations were significantly lower in the 5 and 10% groups （P〈0.01）. In contrast, Cr-Pyr caused a marked increase in the serum nonesterified fatty acid （NEFA）, high-density lipoprotein cholesterol （HDL-C） and creatine kinase （CK） concentrations （P〈0.01）. Supplementation with Cr-Pyr （5 and 10%） in the diet also increased glucagons （GLU）, insulin （INS） or leptin （LEP） contents （P〈0.01）. The expression of hepatic peroxisomal proliferators-activated receptor α （PPAR-α） and carnitine palmitoyl transferase-I （CPT-I）, muscle insulin-like growth factor I （IGF-I） were significantly elevated and myostatin mRNA level was reduced in the 5 and 10% groups （P〈0.05）. It was found that supplementation with 5% Cr-Pyr improves both lipid and protein metabolism by regulating various metabolic parameters of broilers, while not adversely affects growth performance in broiler chickens.

Nitrogen spraying affects seed Bt toxin concentration and yield in Bt cotton

Cotton bolls exhibit the lowest insecticidal efficacy among all organs of Bt cotton,which would ultimately affect the yield formation.The objective of this study was to investigate the effects of different urea concentrations on the seed Bt protein contents,seed cotton yield and the corresponding protein metabolism mechanism.The experiments were conducted during 2017–2018 cotton growing seasons.Two cultivars,Sikang 3(hybrid,SK3)and Sikang 1(conventional,SK1),were treated with six urea concentrations and their seed Bt protein contents were compared during boll formation period.The urea spray concentration had a significant effect on the seed Bt toxin content and seed cotton yield.Spraying of either 5 or 6%urea led to higher insecticidal protein contents and higher seed cotton yield for both cultivars.Moreover,the highest amino acid and soluble protein contents,as well as GPT and GOT activities,and lower protease and peptidase activities were observed at the 5 to 6%urea levels.Significant positive correlations between the seed Bt toxin and amino acid contents,and between the seed Bt toxin content and GPT activities were detected.The lower boll worm number and hazard boll rate were also observed with the 5 to 6%urea treatments,which may be the reason why nitrogen spraying increased the seed cotton yield.Therefore,our results suggested that the seed Bt toxin content and insect resistance were impacted markedly by external nitrogen application,and 5 to 6%urea had the greatest effect on insect resistance.

Alteration of tumor suppressor gene p16 and Rb in gastric cancinogesis

IM To study the alterations of tumor suppressor gene p16 and Rb in the carcinogenesis of the stomach. METHODS Different mucosal biopsies were endoscopically obtained, all samples were immediately fixed with 10% buffered formalin, embedded with paraffin and sectioned serielly. Alterations of p16 and Rb protein in 12 cases of superficial gastritis, 15 atrophic gastritis, 20 atypical hyperplasia and 40 cancerous tissues were detected by the immunohistochemical method (ABC). RESULTS Different degrees of nuclear immunostaining of p16 and Rb occurred on gastric epithelium in different stages of lesions. With the lesions progressing, the positive immunostaining rate of p16 protein had a decreasing tendency (833%→733%→300%→275%), and on the other hand, that of Rb protein had an increasing tendency (250%→467%→600%→675%). A negative correlationship was found between these two parameters in the gastric cancer. Of 40 cases of gastric cancer, a negative relationship was observed in 20 cases. In comparison with both positive (9 cases) and both negative tissues (11 cases), there was a significant difference (500%,225%,275%) (P<005).CONCLUSION Abnormal expression of p16 and Rb plays an important role in gastric carcinogenesis.

Increasing plant density increases Bt toxin concentration of boll wall in cotton by decreasing boll setting speed

Background: In order to uncover the mechanism of significantly reduced insect resistance at the late developmental stage in cotton(Gossypium hirsutum L.),the relationship between boll setting rate under different planting densities and Bacillus thuringiensis(Bt)insecticidal concentrations in the boll wall were investigated in the present study.Two studies were arranged at Yangzhou,China during the 2017–2018 cotton growth seasons.Five planting densities(15000,25000,45000,60000 and 75000 plants per hectare)and the flower-removal treatment were imposed separately on Bt cotton cultivar Sikang3 to arrange different boll setting rates,and the boll setting rates and Bt toxin content were compared.Results: Higher boll setting rate together with lower Bt toxin contents in boll wall was observed under low planting density,whereas lower boll setting rate and higher Bt toxin contents were found under high planting density.Also,higher Bt protein concentration was associated with higher soluble protein content,glutamic-pyruvic transaminase(GPT),and glutamic oxaloacetate transaminase(GOT)activities,but lower amino acid content,and protease and peptidase activities.It was further confirmed that a higher boll setting rate with lower Bt protein content under flower-removal.Conclusions: This study demonstrated that the insecticidal efficacy of boll walls was significantly impacted by boll formation.Reduced protein synthesis and enhanced protein degradation were related to the reduced Bt toxin concentration.

Influence of fever on biliary elements of guinea pigs

IM To study the influence of fever on biliary elements and gallstone formation in guinea pigs. METHODS Sixty guinea pigs were divided into lithogenic (group B) and normal diet (group A) groups, and each group was further divided into control and fever subgroups. The animals in the fever groups were injected subcutaneously with boiled defat milk 1ml/kg, once a week for 4 times to run fever. After 45 days, all the animals were killed. Among them the guinea pigs in the fever groups were injected subcutaneously with turpentine 1ml/kg 36 hours before they were killed, and kept in fever condition. Their gallbladder lumens were examined and bile samples were analyzed. RESULTS The gallstone incidence of the fever group fed with lithogenic diet was the highest, accounting for 40% (6/15). All the animals in fever group had significantly higher concentration of total bile protein and bilirubin than that of the control groups. CONCLUSION The effect of fever on biliary elements plays an important role in the gallstone formation in guinea pigs.

Maternal supplementation with mulberry-leaf flavonoids improves the development of skeletal muscle in the offspring of chickens

The development of skeletal muscle is a crucial factor in determining the meat yield and economic benefits of broiler production.Recent research has shown that mulberry leaves and their extracts can be used to significantly improve the growth performance of livestock and poultry.The present study aims to elucidate the mechanisms involved in the regulation of skeletal muscle development in broiler offspring by dietary mulberry-leaf flavonoids(MLF)supplementation from the perspective of maternal effect theory.A total of 270 Qiling broiler breeder hens were randomly assigned to 3 treatments with different doses of MLF(0,30,60 mg/kg)for 8 weeks before collecting their fertilized eggs.The chicken offspring at 13 and 19 d of embryonic stage,and from 1 to 28 d old after hatching were included in this study.The results showed that maternal supplementation increased the breast muscle weight and body weight of the offspring at the embryo and chick stages(P<0.05).This was followed by increased cross-sectional area of pectoral muscle fibres at 14 d(P<0.05).Further determination revealed a tendency towards increased serum levels of insulin-like growth factor 1(IGF-1)(P=0.092)and muscle fibre count(P=0.167)at 1 d post-hatching following maternal MLF treatment,while serum uric acid(UA)was decreased at 14 d after hatching(P<0.05).Moreover,maternal MLF supplementation significantly up-regulated the mRNA expression of the myogenic regulatory factor Myf5 in skeletal muscle at the both embryonic and growth stages(P<0.05).The relative abundance of the downstream protein of BMPR2,Smad1 and p-Smad1/5/9 in the TGFβsignalling pathway was significantly increased by maternal MLF treatment.Meanwhile,the increased expression of the target protein p-mTOR in the breast muscle of the offspring chicks is in accordance with the improved growth rate of the breast and the body.In conclusion,maternal MLF supplementation can promote muscle protein metabolism and muscle fibre development of chick embryos through upregulation of Myf5 expression and BMP/p-Smad1/5/9 axis,thereby improving growth performance of slow growing broiler.

Primate Torpor: Regulation of Stress-activated Protein Kinases During Daily Torpor in the Gray Mouse Lemur, Microcebus murinus

Very few selected species of primates are known to be capable of entering torpor. This exciting discovery means that the ability to enter a natural state of dormancy is an ancestral trait among primates and, in phylogenetic terms, is very close to the human lineage. To explore the regulatory mechanisms that underlie primate torpor, we analyzed signal transduction cascades to discover those involved in coordinating tissue responses during torpor. The responses of mitogen-activated protein kinase（MAPK） family members to primate torpor were compared in six organs of control（aroused） versus torpid gray mouse lemurs, Microcebus murinus. The proteins examined include extracellular signal-regulated kinases（ERKs）, c-jun NH2-terminal kinases（JNKs）, MAPK kinase（MEK）, and p38, in addition to stress-related proteins p53 and heat shock protein 27（HSP27）. The activation of specific MAPK signal transduction pathways may provide a mechanism to regulate the expression of torpor-responsive genes or the regulation of selected downstream cellular processes. In response to torpor, each MAPK subfamily responded differently during torpor and each showed organ-specific patterns of response. For example, skeletal muscle displayed elevated relative phosphorylation of ERK1/2 during torpor. Interestingly, adipose tissues showed the highest degree of MAPK activation. Brown adipose tissue displayed an activation of ERK1/2 and p38, whereas white adipose tissue showed activation of ERK1/2, p38, MEK, and JNK during torpor. Importantly, both adipose tissues possess specialized functions that are critical for torpor, with brown adipose required for non-shivering thermogenesis and white adipose utilized as the primary source of lipid fuel for torpor. Overall, these data indicate crucial roles of MAPKs in the regulation of primate organs during torpor.